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Abstract：
Agroinfiltration in Nicotiana benthamiana is widely used to transiently express heterologous proteins in plants. However, the state of Agrobacterium itself is not well studied in agroinfiltrated tissues, despite frequent studies of immunity genes conducted through agroinfiltration. Here, we generated a bioluminescent strain of Agrobacterium tumefaciens GV3101 to monitor the luminescence of Agrobacterium during agroinfiltration. By integrating a single copy of the lux operon into the genome, we generated a stable \'AgroLux\' strain, which is bioluminescent without affecting Agrobacterium growth in vitro and in planta. To illustrate its versatility, we used AgroLux to demonstrate that high light intensity post infiltration suppresses both Agrobacterium luminescence and protein expression. We also discovered that AgroLux can detect Avr/Cf-induced immune responses before tissue collapse, establishing a robust and rapid quantitative assay for the hypersensitive response (HR). Thus, AgroLux provides a non-destructive, versatile and easy-to-use imaging tool to monitor both Agrobacterium and plant responses.
摘要：
烟草中的农杆菌浸润被广泛用于在植物中瞬时表达异源蛋白。然而,农杆菌本身的状态在农业浸润组织中没有得到很好的研究，尽管通过农业渗透对免疫基因进行了频繁的研究。这里,我们产生了根癌农杆菌GV3101的生物发光菌株，以监测农杆菌在农杆菌浸润过程中的发光。通过将勒克斯操纵子的单个拷贝整合到基因组中，我们产生了稳定的AgroLux菌株,它是生物发光的，不会影响土壤杆菌在体外和植物中的生长。为了说明它的多功能性，我们使用AgroLux证明浸润后的高光强度抑制了农杆菌发光和蛋白质表达。我们还发现AgroLux可以在组织塌陷之前检测Avr/Cf诱导的免疫反应，建立一个强大的和快速的定量分析超敏反应（HR）。因此，AgroLux提供了一种非破坏性的,多功能和易于使用的成像工具来监测土壤杆菌和植物反应。
